Pyrosequencing analysis reveals changes in intestinal microbiota of healthy adults who received a daily dose of immunomodulatory probiotic strains

Julio Plaza-Díaz, Jose Ángel Fernández-Caballero, Natalia Chueca, Federico García, Carolina Gómez-Llorente, María José Sáez-Lara, Luis Fontana, Ángel Gil, Julio Plaza-Díaz, Jose Ángel Fernández-Caballero, Natalia Chueca, Federico García, Carolina Gómez-Llorente, María José Sáez-Lara, Luis Fontana, Ángel Gil

Abstract

The colon microbiota plays a crucial role in human gastrointestinal health. Current attempts to manipulate the colon microbiota composition are aimed at finding remedies for various diseases. We have recently described the immunomodulatory effects of three probiotic strains (Lactobacillus rhamnosus CNCM I-4036, Lactobacillus paracasei CNCM I-4034, and Bifidobacterium breve CNCM I-4035). The goal of the present study was to analyze the compositions of the fecal microbiota of healthy adults who received one of these strains using high-throughput 16S ribosomal RNA gene sequencing. Bacteroides was the most abundant genus in the groups that received L. rhamnosus CNCM I-4036 or L. paracasei CNCM I-4034. The Shannon indices were significantly increased in these two groups. Our results also revealed a significant increase in the Lactobacillus genus after the intervention with L. rhamnosus CNCM I-4036. The initially different colon microbiota became homogeneous in the subjects who received L. rhamnosus CNCM I-4036. While some orders that were initially present disappeared after the administration of L. rhamnosus CNCM I-4036, other orders, such as Sphingobacteriales, Nitrospirales, Desulfobacterales, Thiotrichales, and Synergistetes, were detected after the intervention. In summary, our results show that the intake of these three bacterial strains induced changes in the colon microbiota.

Trial registration: ClinicalTrials.gov NCT01479543.

Keywords: gut; healthy adults; high-throughput nucleotide sequencing; microbiota; probiotics.

Figures

Figure 1
Figure 1
Compositions of fecal microbiota in 23 healthy adults at the phylum level. The subjects were divided into five groups and received either a placebo, a capsule containing 9 × 109 CFUs of one of the three strains, or a capsule containing 9 × 109 CFUs of a mixture of B. breve CNCM I-4035 and L. rhamnosus CNCM I-4036 each day for 30 days. Each column represents 1 healthy adult, as described in Table 1.
Figure 2
Figure 2
The impacts of L. rhamnosus CNCM I-4036 consumption on the intestinal microbiota. The first washout, intervention and second washout refer to the different L. rhamnosus CNCM I-4036 intake phases. Principal component analysis (PCoA) score plots of the different L. rhamnosus CNCM I-4036 intake phases are shown. The data were compared with those in the Ribosomal Database Project (RDP) using a maximum e-value of 10−5, a minimum identity of 75%, and a minimum alignment length of 15 measured in bp for RNA databases. The data were normalized to values between 0 and 1, and Euclidean distance was measured in the construction of PCoA plots. Each figure represents the composition of the intestinal microbiota of one healthy volunteer.
Figure 3
Figure 3
Tree diagram of the different L. rhamnosus CNCM I-4036 intake phases. (A) First washout. (B) Intervention. (C) Second washout. Each tree diagram represents the composition of the intestinal microbiota of five healthy volunteers who received L. rhamnosus CNCM I-4036. Colors represent orders. FW, First washout; I, intervention; and SW, second washout.
Figure 3
Figure 3
Tree diagram of the different L. rhamnosus CNCM I-4036 intake phases. (A) First washout. (B) Intervention. (C) Second washout. Each tree diagram represents the composition of the intestinal microbiota of five healthy volunteers who received L. rhamnosus CNCM I-4036. Colors represent orders. FW, First washout; I, intervention; and SW, second washout.
Figure 3
Figure 3
Tree diagram of the different L. rhamnosus CNCM I-4036 intake phases. (A) First washout. (B) Intervention. (C) Second washout. Each tree diagram represents the composition of the intestinal microbiota of five healthy volunteers who received L. rhamnosus CNCM I-4036. Colors represent orders. FW, First washout; I, intervention; and SW, second washout.

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